RESUMO
Ophiocordyceps sinensis appears as stroma emerging from underground sclerotium enclosed by the skeleton of Thitarodes moth larvae. However, the actual distribution of the fungus in soil still remains unclarified. In this study, 40 soil samples were used for detection of O. sinensis to confirm its distribution in native habitats using denaturing gradient gel electrophoresis, nested internal transcribed spacer (ITS) PCR, and 454 pyrosequencing methods. The soil samples included six types: Os, where both stromata and host moth larvae were found; NL, representing no signs of stromata, but where moth larvae were found; NOs, where neither stroma nor moth larvae were found; BS, with bare soil without the presence of stroma of O. sinensis or moth larvae; AF, from soil surrounding the stroma; and MP, soil particles firmly wrapping the sclerotium of O. sinensis. Of 40 samples tested, 36 showed positive detection of O. sinensis by at least one of the three detection methods, with positive detection in all six sample types at all five sites. The results showed that traces of O. sinensis can be detected in locations with no macroscopically visible evidence of the fungus or its host and at least 100 m away from such locations.
Assuntos
Cordyceps/fisiologia , Microbiologia do Solo , Animais , China , Cordyceps/química , Cordyceps/genética , DNA Fúngico/química , DNA Fúngico/isolamento & purificação , Eletroforese em Gel de Gradiente Desnaturante , Sequenciamento de Nucleotídeos em Larga Escala , Concentração de Íons de Hidrogênio , Larva/microbiologia , Mariposas/microbiologia , Reação em Cadeia da Polimerase , Solo/química , Solo/classificação , Água/análiseRESUMO
A robust, sensitive, and time-competitive system to detect Candida albicans in less than 30 min in clinical samples based in capped nanoporous anodic alumina (NAA) is developed. In the proposed design, NAA pores are loaded with rhodamine B and then blocked with an oligonucleotide that is able to recognize C. albicans DNA. The capped material shows negligible cargo release, whereas dye delivery is selectively accomplished when genomic DNA from C. albicans is present. This procedure has been successfully applied to detect C. albicans in clinical samples from patients infected with this yeast. When compared with classical C. albicans detection methods, the proposed probe has a short assay time, high sensitivity and selectivity, demonstrating the high potential of this simple design for the diagnosis of infection produced by C. albicans.
Assuntos
Óxido de Alumínio/química , Técnicas Biossensoriais/métodos , Candida albicans/isolamento & purificação , Nanoporos , Oligonucleotídeos/química , Candida albicans/genética , Candida albicans/fisiologia , DNA Fúngico/análise , DNA Fúngico/química , Humanos , Limite de Detecção , Fatores de TempoAssuntos
Ascomicetos/isolamento & purificação , Cromoblastomicose/diagnóstico , Cromoblastomicose/patologia , Joelho/patologia , Adulto , Antifúngicos/administração & dosagem , Técnicas de Tipagem Bacteriana , Cromoblastomicose/tratamento farmacológico , Cromoblastomicose/microbiologia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fungos , Histocitoquímica , Humanos , Hipertermia Induzida , Masculino , Técnicas Microbiológicas , Filogenia , Análise de Sequência de DNA , Pele/patologia , Terbinafina/administração & dosagem , Resultado do TratamentoRESUMO
Species of Coleosporium (Pucciniales) are rust fungi that typically alternate between pines and angiosperms. In North America, species of Coleosporium often infect Solidago (goldenrods), although their taxonomy on these hosts is unresolved. Joseph. C. Arthur and George B. Cummins regarded these as a single species, Coleosporium solidaginis (fide Arthur) or C. asterum (fide Cummins), but later inoculation studies demonstrated the presence of more than one species, distinguishable by their aecial hosts. A more recent taxonomic study of Coleosporium found that specimens on Solidago identified as C. asterum in North America were not conspecific with the type, which is from Japan, prompting the present study. Herein, we conducted a systematic study on ca. 60 collections of Coleosporium infecting species of Asteraceae from North America using regions of ribosomal DNA and morphology of teliospores and basidia. Our data indicate at least three species of Coleosporium occur on Solidago in North America, C. solidaginis, C. montanum comb. nov., which is proposed for the taxon that has commonly been identified as C. asterum in North America, and C. delicatulum, all of which can be differentiated by morphology of their basidia. In addition, the challenges of marker selection for molecular barcoding of rust fungi is discussed.
Assuntos
Basidiomycota/classificação , Basidiomycota/isolamento & purificação , Solidago/microbiologia , Basidiomycota/genética , Basidiomycota/crescimento & desenvolvimento , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Microscopia , América do Norte , Filogenia , Análise de Sequência de DNARESUMO
We analyzed intraspecies genetic variability of the medicinal tinder bracket polypore, Fomes fomentarius, from the Asian part of Russia, including the Ural, Altai, Western Sayan, and Baikal regions. We used nuclear ribosomal DNA internal transcribed spacer (ITS) sequence data as a standard marker for fungal DNA barcoding. In the Asian part of Russia, lineage A occurs as sublineage A2, which differs from sublineage A1 by a single nucleotide insertion at ITS2.3. Sublineage A2 is distributed up to Lake Baikal in the Ural, Altai, and Western Sayan regions. It can be characterized as a Eurasian sublineage of F. fomentarius. Lineage B is also represented by 2 sublineages (B1 and B2), which differ from each other by nucleotide sequences at ITS2.1. Sublineage B1 is represented by a small group of isolates from Asia (Iran, China, Nepal, South Korea), whereas sublineage B2 mainly includes isolates from Europe (Great Britain, Italy, Latvia, Slovakia, Slovenia) and 2 separate samples from Asia (Iran, China); these locales compose the distribution area of F. fomentarius. In the Asian part of Russia, lineage B is represented by sublineage B2 found in the Southern Urals (at the border between Europe and Asia), which is the only area where sublineages A2 and B2 are present. These sublineages are characterized by different substrate spectra: sublineage A2 is predominantly associated with Betula spp. and rarely with Alnus and Larix trees, whereas sublineage B2 does not have a pronounced substrate preference and is found in basidiomes collected from Acer, Duschekia, Prunus, and Salix trees, but not Betula trees. In general, the spectrum of substrates for F. fomentarius lineages A and B in the Asian part of Russia corresponds to that in other parts of this polypore's distribution area. Data are needed on genetic intraspecies variability (polymorphism) in relation to pharmacological properties for further biotechnological cultivation and use of the medicinal fungus F. fomentarius.
Assuntos
Coriolaceae/genética , Variação Genética , Filogenia , Ásia , China , Código de Barras de DNA Taxonômico , DNA Fúngico/química , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , DNA Espaçador Ribossômico/genética , Europa (Continente) , Genótipo , Irã (Geográfico) , Polimorfismo Genético , Federação Russa , Alinhamento de SequênciaRESUMO
The effect of microchip pulsed electric field (MPEF) treatment on lethal and sublethal injury of Pichia rhodanensis (P. rhodanensis) were employed under 100-500 V for 20-100 pulses and the underlying mechanism of MPEF treatment was investigated as well. A 6.48 log10 reduction of P. rhodanensis was achieved at 500V for 80 pulse. The fluorescent staining with Propidium Iodide (PI) verified that the rate of sublethal injury cells maximum up to 27.2% under 200 V. MPEF can cause the damage of cell morphology and ultrastructure, meanwhile causing a decrease in cellular enzymes, antioxidant enzyme activity and cell membrane fluidity. The leakage of intracellular compounds (protein, nucleic acid, K+, Mg2+) and Ca2+-ATPase gradually increased as the growth of voltage, especially the proportion of protein in the supernatants increased from 2.0% to 26.4%. Flow cytometry analysis showed that MPEF has significant effect on membrane potential, but no obvious influence on non-specific esterase. MPEF can cause the changing of the secondary structure of protein, at the same time, double helix structure of DNA became loose and unwinding. These results provide a theoretical guidance for the widespread using of MPEF technology in the application of a non-thermal processing technique for food.
Assuntos
Membrana Celular/ultraestrutura , DNA Fúngico/química , Conservação de Alimentos/instrumentação , Proteínas Fúngicas/química , Pichia/ultraestrutura , ATPases Transportadoras de Cálcio , Membrana Celular/química , Estimulação Elétrica , Eletricidade , Citometria de Fluxo , Conservação de Alimentos/métodos , Potenciais da Membrana , Análise em Microsséries , Conformação Molecular , Pichia/fisiologia , Estrutura Secundária de ProteínaRESUMO
China is home to rich wild and cultivated strains of Lentinus edodes, an important edible and medicinal mushroom. Artificial selection of L. edodes has a long history, and the widely cultivated strains belong to populations different from those of most wild strains. Internal transcribed spacer (ITS) regions have been used as good markers to identify L. edodes populations. But because ITS regions exhibit incomplete concerted evolution, the use of an ITS to identify L. edodes populations has been questioned. The objective of this study was to determine whether the ITS region is suitable for identifying L. edodes populations and which populations the widely cultivated strains and the most wild strains belong to by investigating intraindividual and differential ITS polymorphisms between 44 cultivars and 44 wild strains of L. edodes in China. Intraindividual ITS polymorphism is common in L. edodes strains, and most strains possessed 2 different ITS sequences, which came from their heterokaryons. The genetic polymorphisms of ITS1, 5.8S, and ITS2 in L. edodes strains are distinct. All strains were divided into one 5.8S type (5.8S-A), 2 ITS1 types (ITS1-A and ITS1-B), and 2 ITS2 types (ITS2-A and ITS2-B), which were subdivided into 2 branches (ITS2-A1 and ITS2-A2; ITS2-B1 and ITS2-B2). ITS1/5.8S/ITS2 could be used as a good marker in preliminary classification of L. edodes strains in China. It not only exhibited classified information of ITS1, 5.8S, and ITS2 for each strain at the same time, it also indicated whether the strain was heterozygous. The 44 cultivated strains were mainly the A/A/A1 type, and the 44 wild strains were mainly the A/A/A2 and other mixed types.
Assuntos
DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Variação Genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Cogumelos Shiitake/classificação , Cogumelos Shiitake/genética , China , Análise por Conglomerados , DNA Fúngico/química , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , Genótipo , Filogenia , Cogumelos Shiitake/isolamento & purificaçãoRESUMO
Global climate and land use change are altering plant and soil microbial communities worldwide, particularly in arctic and alpine biomes where warming is accelerated. The widespread expansion of woody shrubs into historically herbaceous alpine plant zones is likely to interact with climate to affect soil microbial community structure and function; however, our understanding of alpine soil ecology remains limited. This study aimed to (i) determine whether the diversity and community composition of soil fungi vary across elevation gradients and to (ii) assess the impact of woody shrub expansion on these patterns. In the White Mountains of California, sagebrush (Artemisia rothrockii) shrubs have been expanding upwards into alpine areas since 1960. In this study, we combined observational field data with a manipulative shrub removal experiment along an elevation transect of alpine shrub expansion. We utilized next-generation sequencing of the ITS1 region for fungi and joint distribution modelling to tease apart effects of the environment and intracommunity interactions on soil fungi. We found that soil fungal diversity declines and community composition changes with increasing elevation. Both abiotic factors (primarily soil moisture and soil organic C) and woody sagebrush range expansion had significant effects on these patterns. However, fungal diversity and relative abundance had high spatial variation, overwhelming the predictive power of vegetation type, elevation and abiotic soil conditions at the landscape scale. Finally, we observed positive and negative associations among fungal taxa which may be important in structuring community responses to global change.
Assuntos
Artemisia/fisiologia , Fungos/genética , Micobioma , Microbiologia do Solo , Altitude , Artemisia/genética , Biodiversidade , California , Mudança Climática , DNA Fúngico/química , Variação Genética , Análise de Sequência de DNARESUMO
During homologous recombination, Rad51 forms a nucleoprotein filament with single-stranded DNA (ssDNA) that undergoes strand exchange with homologous double-stranded DNA (dsDNA). Here, we use real-time analysis to show that strand exchange by fission yeast Rad51 proceeds via two distinct three-strand intermediates, C1 and C2. Both intermediates contain Rad51, but whereas the donor duplex remains intact in C1, the ssDNA strand is intertwined with the complementary strand of the donor duplex in C2. Swi5-Sfr1, an evolutionarily conserved recombination activator, facilitates the C1-C2 transition and subsequent ssDNA release from C2 to complete strand exchange in an ATP-hydrolysis-dependent manner. In contrast, Ca2+, which activates the Rad51 filament by curbing ATP hydrolysis, facilitates the C1-C2 transition but does not promote strand exchange. These results reveal that Swi5-Sfr1 and Ca2+ have different activation modes in the late synaptic phase, despite their common function in stabilizing the presynaptic filament.
Assuntos
Dano ao DNA , DNA de Cadeia Simples , Nucleoproteínas/química , Rad51 Recombinase/química , Proteínas de Schizosaccharomyces pombe/química , Schizosaccharomyces/química , Trifosfato de Adenosina/química , Cálcio/química , Simulação por Computador , DNA Fúngico/química , Fluorometria , Recombinação Homóloga , Hidrólise , Íons , Cinética , Ligação Proteica , Domínios Proteicos , Análise de Regressão , Proteínas de Schizosaccharomyces pombe/metabolismoRESUMO
BACKGROUND: Potato virus Y (PVY) is one of the most economically important pathogen of potato that is present as biologically distinct strains. The virus-derived small interfering RNAs (vsiRNAs) from potato cv. Russet Burbank individually infected with PVY-N, PVY-NTN and PVY-O strains were recently characterized. Plant defense RNA-silencing mechanisms deployed against viruses produce vsiRNAs to degrade homologous viral transcripts. Based on sequence complementarity, the vsiRNAs can potentially degrade host RNA transcripts raising the prospect of vsiRNAs as pathogenicity determinants in virus-host interactions. This study investigated the global effects of PVY vsiRNAs on the host potato transcriptome. METHODS: The strain-specific vsiRNAs of PVY, expressed in high copy number, were analyzed in silico for their proclivity to target potato coding and non-coding RNAs using psRobot and psRNATarget algorithms. Functional annotation of target coding transcripts was carried out to predict physiological effects of the vsiRNAs on the potato cv. Russet Burbank. The downregulation of selected target coding transcripts was further validated using qRT-PCR. RESULTS: The vsiRNAs derived from biologically distinct strains of PVY displayed diversity in terms of absolute number, copy number and hotspots for siRNAs on their respective genomes. The vsiRNAs populations were derived with a high frequency from 6 K1, P1 and Hc-Pro for PVY-N, P1, Hc-Pro and P3 for PVY-NTN, and P1, 3' UTR and NIa for PVY-O genomic regions. The number of vsiRNAs that displayed interaction with potato coding transcripts and number of putative coding target transcripts were comparable between PVY-N and PVY-O, and were relatively higher for PVY-NTN. The most abundant target non-coding RNA transcripts for the strain specific PVY-derived vsiRNAs were found to be MIR821, 28S rRNA,18S rRNA, snoR71, tRNA-Met and U5. Functional annotation and qRT-PCR validation suggested that the vsiRNAs target genes involved in plant hormone signaling, genetic information processing, plant-pathogen interactions, plant defense and stress response processes in potato. CONCLUSIONS: The findings suggested that the PVY-derived vsiRNAs could act as a pathogenicity determinant and as a counter-defense strategy to host RNA silencing in PVY-potato interactions. The broad range of host genes targeted by PVY vsiRNAs in infected potato suggests a diverse role for vsiRNAs that includes suppression of host stress responses and developmental processes. The interactome scenario is the first report on the interaction between one of the most important Potyvirus genome-derived siRNAs and the potato transcripts.
Assuntos
Interações Hospedeiro-Patógeno , Doenças das Plantas/virologia , Potyvirus/patogenicidade , RNA de Plantas/análise , RNA Interferente Pequeno/metabolismo , RNA Viral/metabolismo , Solanum tuberosum/virologia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA de Plantas/química , DNA de Plantas/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Perfilação da Expressão Gênica , Filogenia , Potyvirus/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética , RNA Viral/genética , Análise de Sequência de DNARESUMO
"Paracoccidioidomycosis ceti" is a rare zoonotic fungal infection affecting dolphins and is endemic worldwide. The causative agents are Paracoccidioides species; however, it is impossible to isolate the fungal species. We isolated Trichosporon asteroides from multifocal, irregularly raised skin lesions on a female bottlenose dolphin (Tursiops truncatus) captured off coast of Japan, which was suspected to have "paracoccidioidomycosis ceti." An abundance of round, yeast-like cells was detected in a potassium hydroxide direct-mount specimen of the skin samples; however, nested PCR targeting the partial sequence of 43-kDa glycoprotein-coding gene correspondent to Paracoccidioides sp. was negative. Biopsied tissue samples were cultured on brain heart infusion agar plates supplemented with chloramphenicol, 1% yeast extract, and 4% sodium chloride (4% NaCl-BHI), on Mycosel agar with 4% sodium chloride (4% NaCl-Mycosel), and on potato dextrose agar supplemented with chloramphenicol (CPDA) at 35 °C for 4 weeks. Cream-colored and wrinkled colonies consisting of hyphae and arthroconidia grew on 4% NaCl-BHI and CPDA, while film-like colonies composed of arthroconidia and round yeast-like cells developed on 4% NaCl-Mycosel. Although these primary cultures resembled fresh isolates of P. brasiliensis, they were identified as Trichosporon asteroides based on routine mycological studies and the internal transcribed spacer regions of ribosomal RNA sequences. The results suggested that trichosporonosis caused by T. asteroides might remain latent among cases of "paracoccidioidomycosis ceti" diagnosed without cultures and molecular biological analysis.
Assuntos
Golfinho Nariz-de-Garrafa , Dermatomicoses/veterinária , Trichosporon/classificação , Trichosporon/isolamento & purificação , Tricosporonose/veterinária , Animais , Biópsia , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Dermatomicoses/diagnóstico , Dermatomicoses/patologia , Feminino , Japão , Técnicas de Tipagem Micológica , Filogenia , Análise de Sequência de DNA , Trichosporon/genética , Tricosporonose/diagnóstico , Tricosporonose/patologiaRESUMO
Auricularia auricula-judae is an edible and medicinal fungus ranking fourth in production among the edible fungi cultivated worldwide. White villous disease is rampant in Northeast China; it infects the fruiting bodies of A. auricula-judae by forming a white mycelial layer on its ventral side. The disease not only causes an unacceptable morphological appearance and a poor-quality product, but it also significantly reduces the yield. In this study, based on fungal morphology, ribosomal DNA internal transcribed spacer sequences, identification of species-specific primers, and the pathogenicity of the mycelia and spores, 2 fungal pathogens were isolated and identified as Fusarium equiseti and F. sporotrichioides.
Assuntos
Agaricales/crescimento & desenvolvimento , Basidiomycota/crescimento & desenvolvimento , Carpóforos/crescimento & desenvolvimento , Fusarium/classificação , Fusarium/isolamento & purificação , China , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fusarium/genética , Fusarium/crescimento & desenvolvimento , Filogenia , Análise de Sequência de DNARESUMO
The aim of this study was to characterize 3 wild culinary-medicinal mushrooms using molecular tools and to analyze their antioxidant activity. Antioxidant properties were studied by evaluating free radical scavenging, reducing power, and chelating effect. The mushrooms were identified as Lentinus squarrosulus, L. tuber-regium, and Macrocybe gigantean by amplifying internal transcribed spacer regions of ribosomal DNA. The results demonstrated that the methanolic extract of M. gigantean has the highest free radical scavenging effect and chelating effect, whereas the methanolic extract of L. squarrosulus has the highest reducing power. The highest total phenol content and the most ascorbic acid were found in the M. gigantean extracts. Among the 3 mushroom extracts, M. gigantean displayed the most potent antioxidant activity. Molecular characterization using the nuclear ribosomal internal transcribed spacer region as a universal DNA marker was an effective tool in the identification and phylogenetic analysis of the studied mushrooms. The study also indicated that these wild macrofungi are rich sources of natural antioxidants.
Assuntos
Agaricales/química , Antioxidantes/análise , Sequestradores de Radicais Livres/análise , Agaricales/classificação , Agaricales/genética , Ácido Ascórbico/análise , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Índia , Fenóis/análise , Filogenia , Análise de Sequência de DNARESUMO
Soil microbes play important roles in plant growth and health. Little is known about the differences of soil microbes between healthy and bacterial wilt infected soils with Ralstonia solanacearum. By Illumina-MiSeq sequencing of 16S rRNA and 18S rRNA gene amplicons, we found the soil microbial composition and diversity were distinct between healthy and bacterial wilt infected soils. Soil microbial community varied at different plant growth stages due to changes of root exudates composition and soil pH. Healthy soils exhibited higher microbial diversity than the bacterial wilt infected soils. More abundant beneficial microbes including Bacillus, Agromyces, Micromonospora, Pseudonocardia, Acremonium, Lysobacter, Mesorhizobium, Microvirga, Bradyrhizobium, Acremonium and Chaetomium were found in the healthy soils rather than the bacterial wilt infected soils. Compared to bacterial wilt infected soils, the activities of catalase, invertase and urease, as well as soil pH, available phosphorous and potassium content, were all significantly increased in the healthy soils. In a conclusion, the higher abundance of beneficial microbes are positively related the higher soil quality, including better plant growth, lower disease incidence, and higher nutrient contents, soil enzyme activities and soil pH.
Assuntos
Bactérias/isolamento & purificação , Biota , Fungos/isolamento & purificação , Doenças das Plantas/microbiologia , Microbiologia do Solo , Bactérias/classificação , Bactérias/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Fungos/classificação , Fungos/genética , Concentração de Íons de Hidrogênio , Metagenômica , Fósforo/análise , Filogenia , Potássio/análise , RNA Ribossômico 16S/genética , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Solo/químicaRESUMO
Alternaria species, primarily the small-spored Alternaria alternata and the large-spored Alternaria solani, are considered a serious threat to potato cultivation. To develop control strategies, it is important to gain insight into the Alternaria population. Based on the sequence analyses of the internal transcribed spacer region (ITS) and the glyceraldehyde-3-phosphate dehydrogenase gene, the small-spored and large-spored Alternaria isolates could be separated from each other. Sequence analyses of the calmodulin gene and the RNA polymerase second largest subunit showed that besides A. solani also A. grandis and A. protenta were present in the large-spored Alternaria population. Sequence analyses of the Alternaria major allergen gene Alt a 1 and the elongation factor-α revealed that both A. alternata and species belonging to the Alternaria arborescens species complex were present in the small-spored Alternaria population. Furthermore, according to the histone h3 sequence the members of the A. arborescens species complex could be subdivided into two groups. Concerning the fitness, it was concluded that the mycelium growth rate of the large-spored isolates was significantly lower compared to the growth rate of the small-spored isolates. In contrast, the spore-germinating capacity and early growth of the large-spored isolates was greater compared to those of the small-spored isolates. Within the groups of small-spored and large-spored isolates there were no significant differences in fitness between the species.
Assuntos
Alternaria/classificação , Alternaria/isolamento & purificação , Solanum tuberosum/microbiologia , Alternaria/genética , Alternaria/crescimento & desenvolvimento , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Europa (Continente) , Proteínas Fúngicas/genética , Micélio/crescimento & desenvolvimento , Filogenia , Análise de Sequência de DNA , Esporos Fúngicos/crescimento & desenvolvimentoRESUMO
Filamentous fungi are efficient producers of lipases. The present study focuses on identification of a potent lipolytic fungus and enhancement of lipase production through optimization of nutritional and cultural conditions under submerged fermentation. Molecular characterization of the fungus by 18S rDNA sequencing revealed its identity as Aspergillus tamarii with 98% homology. Maximum lipase production was noted in mineral salts medium supplemented with coconut oil (2.5%, v/v). A combination of ammonium chloride (2%, w/v) and tryptone (2%, w/v) facilitated maximum lipase production at pH 5 of the production medium. A carbon: nitrogen ratio of 1:4 led to significant (p < 0.00008) increase in the enzyme production in the presence of surfactant cetyltrimethylammonium bromide (0.5%, w/v). Maximum lipase activity (2,32,500 ± 192 U/ml/min) was recorded after 7 days of incubation at 25 °C on a rotary shaker at 120 rpm. A 9.8-fold increase in lipase activity was recorded after optimization of the process parameters. Addition of crude lipase enhanced the oil stain removal activity of a commercially available detergent by 2.2-fold. The current findings suggest the potentiality of this fungal lipase to be used in detergent formulation.
Assuntos
Aspergillus/enzimologia , Aspergillus/metabolismo , Lipase/biossíntese , Óleos de Plantas/metabolismo , Aspergillus/crescimento & desenvolvimento , Óleo de Coco , Meios de Cultura/química , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Inibidores Enzimáticos/metabolismo , Fermentação , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Tensoativos/metabolismo , TemperaturaRESUMO
Ophiocordyceps sinensis is a valuable traditional Chinese medicine with a high market price. In this study, a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) method based on 2 enzymes was developed to distinguish O. sinensis from 6 common counterfeit species. To verify the applicability of this method, we experimentally tested O. sinensis organisms, tablet preparations made from O. sinensis, and cultured mycelia isolated from O. sinensis. To validate the results from this PCR-RFLP method, all real samples were identified by internal transcribed spacer sequencing. This is, to our knowledge, the first time the PCR-RFLP method has been applied to identify O. sinensis. The selection of 2 restrictive enzymes for identification dramatically improved the accuracy and efficiency of this method. It is the great advantage of this method that sampling from either of 2 parts of O. sinensis-the fruiting body or the caterpillar body-would not cause any difference in the final experimental results. Therefore, this method is not only feasible for testing crude drugs of O. sinensis but it is also useful when the crude drugs are broken down into powder or made into tablets, demonstrating the promising prospect of application in quality control.
Assuntos
Ascomicetos/classificação , Ascomicetos/genética , Medicamentos Falsificados/isolamento & purificação , Medicina Tradicional Chinesa/normas , Medicamentos Falsificados/classificação , Medicamentos Falsificados/economia , DNA Fúngico/química , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , DNA Espaçador Ribossômico/genética , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Controle de QualidadeRESUMO
An updated overview of the outcome of studies conducted on the culinary-medicinal mushroom Pleurotus nebrodensis is presented by placing emphasis on the clarification of the taxonomic identity of P. nebrodensis and other related taxa possessing entirely white to cream basidiomes, which grow in association with different plants of the family Apiaceae. Cultivation techniques, quality of the product sold and sales price, as well as nutritional and medicinal aspects are discussed. Taking also into consideration the high economic importance of P. nebrodensis, it is essential to proceed with the verification of the commercial strains currently available in the international market under the name of "P. nebrodensis" since it is very probable that many (or most) of them do not represent the real P. nebrodensis. TO confirm this hypothesis, an in silico analysis was conducted on a large of number of ITS1-5.8S-ITS2 rRNA sequences deposited in the National Center for Biotechnology Information database under the name P. nebrodensis. Results demonstrated that all "P nebrodensis" material examined from China (plus several sequences of no reported origin) corresponded to P. eryngii subsp. tuoliensis, with only 2 exceptions, which were grouped within P. eryngii sensu stricto. The real P. nebrodensis biological material from Italy and Greece is certified and is available upon request by the authors at the University of Palermo and the Agricultural University of Athens.
Assuntos
Produtos Biológicos/economia , Alimentos/economia , Pleurotus/classificação , Produtos Biológicos/normas , China , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Alimentos/normas , Geografia , Grécia , Humanos , Itália , Filogenia , Pleurotus/química , Pleurotus/genética , Pleurotus/crescimento & desenvolvimento , Especificidade da EspécieRESUMO
Polyporoid and lentinoid fungi contain the important producers of substances having immunomodulatory, antitumoral, antiviral, and antihyperlipidemic effects. The discovery of several phylogenetic lines within the lentinoid-polyporoid continuum will help with target metabolomic analysis of species still not studied in pharmacological respects. The purpose of the present work was to increase a resolution in the lentinoid-polyporoid phylogenetic zone by means of selection of both the main representatives of Lentinus-related genera and poorly known/intermediate taxa such as Lentinus suavissimus, Neofavolus spp., and the resupinate part of Polyporus (genera Perenniporia and Pachykytospora) in the context of the basic structure of the Polyporales tree. The molecular phylogeny of highlighting all the polyporoid and lentinoid nodes was reconstructed using nLSU ITS rDNA and TEF datasets. The data obtained from ITS, TEF, and LSU coincide in support of core Polyporaceae of 10 clades corresponded to the generic level and 7 of these (Cerioporus, Cladomeris, Favolus, Lentinus, Neofavolus, Picipes, and Polyporus s.str.) contain generic units characterized by polyporoid or lentinoid morphotypes. The other 2 clades containing lentinoid taxa are outside the core Polyporaceae, namely Panus (Meruliaceae, Polyporales) and Neolentinus (Gloeophyllaceae, Gloeophyllales). A new genus, Picipes, is described and 25 new combinations are proposed.
Assuntos
Lentinula/classificação , Polyporaceae/classificação , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Proteínas Fúngicas/genética , Lentinula/química , Lentinula/genética , Filogenia , Polyporaceae/química , Polyporaceae/genética , Polyporus/química , Polyporus/classificação , Polyporus/genética , Análise de Sequência de DNARESUMO
Fusisporium solani was described as the causal agent of a dry rot of potato in Germany in the mid 19th century. As Fusarium solani, the species became known as a plurivorous plant pathogen, endophyte, decomposer, and opportunistic pathogen of humans and nutritional symbiont of insects. In parallel, it became evident that the morphologically defined species F. solani represents a phylogenetically and biologically complex group of often morphologically cryptic species that has come to be known in part as the F. solani species complex (FSSC), accommodating several formae speciales and mating populations/biological species. The FSSC currently includes more than 60 phylogenetic species. Several of these have been named, but the majority remains unnamed and the identity of F. solani sensu stricto is unclear. To promote further taxonomic developments in the FSSC, lectoand epitypification is proposed for Fusisporium solani Although no type material for F. solani is known to exist, the species was abundantly illustrated in the protologue. Thus, a relevant illustration provided by von Martius is selected as the lectotype. The epitype selected here originates from a rotting potato collected in a field in Slovenia. This strain causes a dry rot of artificially inoculated potatoes. It groups in the heretofore unnamed phylogenetic species 5, which is nested within clade 3 of the FSSC (FSSC 5). Members of this phylogenetic species have a wide geographic distribution and include soil saprotrophs and plant and opportunistic human pathogens. This typification is consistent with the original description of Fusisporium solani and the concept of F. solani as a widely distributed soil inhabitant and pathogen.